A protocol for isolation and visualization of yeast nuclei by scanning electron microscopy
作者： Stephen Murray , Elena Kiseleva
DOI: 10.1016/S0091-679X(08)00419-6
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摘要: This article describes a protocol that details methods for the isolation of yeast nuclei from budding (Saccharomyces cerevisiae) and fission (Schizosaccharomyces pombe), immunogold labelling proteins, visualization by Field Emission Scanning Electron Microscopy (FESEM). involves removal cell wall nucleus within, followed subsequent processing high resolution microscopy. The nuclear step is performed enzymatic treatment cells to rupture generate spheroplasts (cells have partially lost their characteristic shape), centrifugation. has been optimized envelope (NE), pore complexes (NPCs), associated cytoskeletal structures. Samples, once processed FESEM, can be stored under vacuum weeks, allowing considerable time image acquisition.
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Th.H. Rozijn, G.J.M. Tonino, Studies on the yeast nucleus Biochimica et Biophysica Acta (BBA) - Specialized Section on Nucleic Acids and Related Subjects. ,vol. 91, pp. 105- 112 ,(1964) , 10.1016/0926-6550(64)90174-4
Paul Nurse, Yvonne Bissett, Gene required in G1 for commitment to cell cycle and in G2 for control of mitosis in fission yeast. Nature. ,vol. 292, pp. 558- 560 ,(1981) , 10.1038/292558A0
T. G. BONNEY, Mountaineering in the Sierra Nevada Nature. ,vol. 6, pp. 78- 78 ,(1872) , 10.1038/006078A0
W.W. Sillevis Smitt, G. Nanni, Th.H. Rozijn, G.J.M. Toning, Sedimentation characteristics of RNA from isolated yeast nuclei Experimental Cell Research. ,vol. 59, pp. 440- 446 ,(1970) , 10.1016/0014-4827(70)90651-8
R. S. S. Fraser, Paul Nurse, Novel cell cycle control of RNA synthesis in yeast. Nature. ,vol. 271, pp. 726- 730 ,(1978) , 10.1038/271726A0
Paul Nurse, Cell divison: Coordinating histone transcription and DNA replication Nature. ,vol. 302, pp. 378- 378 ,(1983) , 10.1038/302378A0
Jurgen A.W. Heymann, Mike Hayles, Ingo Gestmann, Lucille A. Giannuzzi, Ben Lich, Sriram Subramaniam, Site-specific 3D imaging of cells and tissues with a dual beam microscope. Journal of Structural Biology. ,vol. 155, pp. 63- 73 ,(2006) , 10.1016/J.JSB.2006.03.006
Albert K. Ho, Tian Xiang Shen, Kathryn J. Ryan, Elena Kiseleva, Marilyn Aach Levy, Terence D. Allen, Susan R. Wente, Assembly and preferential localization of Nup116p on the cytoplasmic face of the nuclear pore complex by interaction with Nup82p. Molecular and Cellular Biology. ,vol. 20, pp. 5736- 5748 ,(2000) , 10.1128/MCB.20.15.5736-5748.2000
Jerry Lynn Allen, Michael G. Douglas, Organization of the nuclear pore complex in Saccharomyces cerevisiae. Journal of Ultrastructure and Molecular Structure Research. ,vol. 102, pp. 95- 108 ,(1989) , 10.1016/0889-1605(89)90047-5
Terence D Allen, Sandra A Rutherford, S Murray, Helen S Sanderson, Fiona Gardiner, Elena Kiseleva, Martin W Goldberg, Sheona P Drummond, None, A protocol for isolating Xenopus oocyte nuclear envelope for visualization and characterization by scanning electron microscopy (SEM) or transmission electron microscopy (TEM). Nature Protocols. ,vol. 2, pp. 1166- 1172 ,(2007) , 10.1038/NPROT.2007.137